Aron Gyorgypal, Antash Chaturvedi, Viki Chopda, Haoran Zhang, Shishir P S Chundawat
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引用次数: 0
Abstract
The choice of media and feeds significantly influences the performance of Chinese Hamster Ovary (CHO) mammalian cell cultures in producing desired biologics like monoclonal antibodies (mAb). Sub-optimal nutrient feed/media composition can severely impact cell proliferation and the quality of the final mAb product. For instance, proper protein glycosylation, crucial for mAb stability, safety, and efficacy, heavily relies on cell culture conditions. Currently, starter CHO culture media and daily supplemental feeds used in industrial manufacturing consist of proprietary composition of nutrients critical for mAb production. Standardized optimal media/feed combinations necessary for different cell lines are often lacking, necessitating individualized optimization for each cell line and mAb product. Here, we focused on a CHO-K1 cell line engineered to produce a Trastuzumab biosimilar and evaluated the effects of fourteen commercially relevant basal media and seven feeds on cell culture parameters such as viable cell density, viability, nutrient consumption, metabolite production, mAb titer, and mAb N-glycosylation. Our findings demonstrate clearly that the compositions of the basal medium and feed play a pivotal role in enhancing cell growth and mAb production. This work offers valuable insights into strategies for optimizing feed/media composition for glycosylated monoclonal antibody production using CHO cells.
Supplementary information: The online version contains supplementary material available at 10.1007/s10616-024-00690-7.
培养基和饲料的选择在很大程度上影响着中国仓鼠卵巢(CHO)哺乳动物细胞培养在生产单克隆抗体(mAb)等所需生物制品方面的性能。营养饲料/培养基组成不合理会严重影响细胞增殖和最终 mAb 产品的质量。例如,适当的蛋白质糖基化对 mAb 的稳定性、安全性和有效性至关重要,在很大程度上依赖于细胞培养条件。目前,工业生产中使用的起始 CHO 培养基和日常补充饲料由对 mAb 生产至关重要的专有营养成分组成。不同细胞系所需的标准化最佳培养基/饲料组合往往缺乏,因此需要针对每个细胞系和 mAb 产品进行个性化优化。在这里,我们重点研究了为生产曲妥珠单抗生物类似物而设计的 CHO-K1 细胞系,并评估了 14 种商业相关的基础培养基和 7 种饲料对细胞培养参数的影响,如存活细胞密度、存活率、营养消耗、代谢产物产量、mAb 滴度和 mAb N-糖基化。我们的研究结果清楚地表明,基础培养基和饲料的成分在提高细胞生长和 mAb 产量方面起着关键作用。这项工作为利用 CHO 细胞生产糖基化单克隆抗体的饲料/培养基成分优化策略提供了有价值的见解:在线版本包含补充材料,可查阅 10.1007/s10616-024-00690-7。
期刊介绍:
The scope of the Journal includes:
1. The derivation, genetic modification and characterization of cell lines, genetic and phenotypic regulation, control of cellular metabolism, cell physiology and biochemistry related to cell function, performance and expression of cell products.
2. Cell culture techniques, substrates, environmental requirements and optimization, cloning, hybridization and molecular biology, including genomic and proteomic tools.
3. Cell culture systems, processes, reactors, scale-up, and industrial production. Descriptions of the design or construction of equipment, media or quality control procedures, that are ancillary to cellular research.
4. The application of animal/human cells in research in the field of stem cell research including maintenance of stemness, differentiation, genetics, and senescence, cancer research, research in immunology, as well as applications in tissue engineering and gene therapy.
5. The use of cell cultures as a substrate for bioassays, biomedical applications and in particular as a replacement for animal models.